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Identification of protein targets for the antidepressant effects of Kai-Xin-San in Chinese medicine using isobaric tags for relative and absolute quantitation

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Neural Regeneration Research 2020年第2期15卷 302-310页

作者： Xian-Zhe Dong Dong-Xiao Wang Tian-Yi Zhang Xu Liu Ping Liu Yuan Hu Department of Clinical Pharmacology and Pharmacy Center of PharmacyChinese PLA General HospitalBeijingChina Department of Pharmacy Xuanwu Hospital of Capital Medical UniversityBeijingChina

Kai-Xin-San consists of Ginseng Radix, Polygalae Radix, Acori Tatarinowii Rhizoma, and Poria at a ratio of 3:3:2:2. Kai-Xin-San has been widely used for the treatment of emotional disorders in China. However, no studies have identified the key proteins implicated in response to Kai-Xin-San treatment. In this study, rat models of chronic mild stress were established using different stress methods over 28 days. After 14 days of stress stimulation, rats received daily intragastric administrations of 600 mg/kg Kai-Xin-San. The sucrose preference test was used to determine depression-like behavior in rats, while isobaric tags were used for relative and absolute quantitation-based proteomics to identify altered proteins following Kai-Xin-San treatment. Kai-Xin-San treatment for 2 weeks noticeably improved depression-like behaviors in rats with chronic mild stress. We identified 33 differentially expressed proteins: 7 were upregulated and 26 were downregulated. Functional analysis showed that these differentially expressed proteins participate in synaptic plasticity, neurodevelopment, and neurogenesis. Our results indicate that Kai-Xin-San has an important role in regulating the key node proteins in the synaptic signaling network, and are helpful to better understand the mechanism of the antidepressive effects of Kai-Xin-San and to provide objective theoretical support for its clinical application. The study was approved by the Ethics Committee for Animal Research from the Chinese PLA General Hospital(approval No. X5-2016-07) on March 5, 2016.

关键词： brain-derived neurotrophic factor signal pathway depression isobaric tags for relative and absolute quantitation Kai-Xin-San neurogenesis protein network proteomics analysis synaptic plasticity traditional Chinese medicine

Different protein expression patterns in rat spinal nerves during wallerian degeneration assessed using isobaric tags for relative and absolute quantitation proteomics profiling

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Neural Regeneration Research 2020年第2期15卷 315-323页

作者： Shuai Wei Xue-Zhen Liang Qian Hu Wei-Shan Wang Wen-Jing Xu Xiao-Qing Cheng Jiang Peng Quan-Yi Guo Shu-Yun Liu Wen Jiang Xiao Ding Gong-Hai Han Ping Liu Chen-Hui Shi Yu Wang The First Affiliated Hospital of Medical College Shihezi UniversityShiheziXinjiang Uygur Autonomous RegionChina Institute of Orthopedics Chinese PLA General HospitalBeijingChina Co-Innovation Center of Neuroregeneration Nantong UniversityNantongJiangsu ProvinceChina The First Clinical Medical School Shandong University of Traditional Chinese MedicineJinanShandong ProvinceChina Kunming Medical University KunmingYunnan ProvinceChina Shanxi Medical University TaiyuanShanxi ProvinceChina

Sensory and motor nerve fibers of peripheral nerves have different anatomies and regeneration functions after injury. To gain a clear understanding of the biological processes behind these differences, we used a labeling technique termed isobaric tags for relative and absolute quantitation to investigate the protein profiles of spinal nerve tissues from Sprague-Dawley rats. In response to Wallerian degeneration, a total of 626 proteins were screened in sensory nerves, of which 368 were upregulated and 258 were downregulated. In addition, 637 proteins were screened in motor nerves, of which 372 were upregulated and 265 were downregulated. All identified proteins were analyzed using the Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analysis of bioinformatics, and the presence of several key proteins closely related to Wallerian degeneration were tested and verified using quantitative real-time polymerase chain reaction analyses. The differentially expressed proteins only identified in the sensory nerves were mainly relevant to various biological processes that included cell-cell adhesion, carbohydrate metabolic processes and cell adhesion, whereas differentially expressed proteins only identified in the motor nerves were mainly relevant to biological processes associated with the glycolytic process, cell redox homeostasis, and protein folding. In the aspect of the cellular component, the differentially expressed proteins in the sensory and motor nerves were commonly related to extracellular exosomes, the myelin sheath, and focal adhesion. According to the Kyoto Encyclopedia of Genes and Genomes, the differentially expressed proteins identified are primarily related to various types of metabolic pathways. In conclusion, the present study screened differentially expressed proteins to reveal more about the differences and similarities between sensory and motor nerves during Wallerian degeneration. The present findings could provide a reference point for a fut

关键词： gene ontology Kyoto Encyclopedia of Genes and Genomes isobaric tags for relative and absolute quantitation motor nerve proteomics sensory nerve spinal nerve Wallerian degeneration

Preparation and Application of Silica-based Perfusion Packing of Size Exclusion Chromatography for quantitation of Polyacrylamide in Enhanced Oil Recovery Systems

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Petroleum Science 2007年第2期4卷 82-90页

作者： Sui Xihua Zha Qingfang Wu Mingbo Guo Yansheng State Key Laboratory of Heavy Oil Processing China University of Petroleum （East China） Dongying Shandong 257061 China

The synthesis and characterization of a novel macroporous silica derived size exclusion chromatography （SEC） packing for quantitative analysis of high molecular weight （MW） polyacrylamide （PAM） are presented. Using this packing, a fast, sensitive and reproducible approach for quantitation of super high-MW PAM in demanding enhanced oil recovery （EOR） waters was developed and the effect of synthesis parameters on the properties of resultant materials was investigated. These parameters include salt addition, reaction temperature and duration, activation condition of functional groups on the silica surface, as well as the reaction cycles required for optimal silica modification. Moreover, SEC analysis conditions, such as mobile phase composition, flow rate, detection and sample preparation, were also explored and an optimal analysis protocol was developed. Under this optimized SEC analysis conditions, the synthesized macroporous materials proved satisfactory for quantification of PAM with average MW up to 22 million Daltons. An SEC analysis required less than few minutes with a detection limit of 1 ng, a linear response range of 0.1 to 75 mg/L with squared R value of 0.99 and reproducibility better than 9.2% RSD （relative standard deviation）. The analysis of PAM in highly saline oilfield production water containing interfering high MW polymeric surfactants indicated the recovery ranges from 92.5% to 110.1% for 1.0 mg/L PAM and 94.2% to 103.8% for 50 mg/L PAM solution. This study presented for the ftrst time that the reliable quantization of high MW PAM in highly demanding EOR waters can be achieved by SEC.

关键词： Macroporous silica size exclusion chromatography quantitation polyacrylamide

How to use open-pFind in deep proteomics data analysis?—A protocol for rigorous identification and quantitation of peptides and proteins from mass spectrometry data

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Biophysics Reports 2021年第3期7卷 207-226页

作者： Guangcan Shao Yong Cao Zhenlin Chen Chao Liu Shangtong Li Hao Chi Meng-Qiu Dong School of Life Sciences Peking University National Institute of Biological Sciences Beijing Tsinghua Institute of Multidisciplinary Biomedical Research Tsinghua University Key Lab of Intelligent Information Processing of Chinese Academy of Sciences (CAS) University of CASInstitute of Computing TechnologyCAS University of Chinese Academy of Sciences Beijing Advanced Innovation Center for Big Data-Based Precision Medicine School of Medicine and EngineeringBeihang University

High-throughput proteomics based on mass spectrometry(MS) analysis has permeated biomedical science and propelled numerous research projects. p Find 3 is a database search engine for high-speed and in-depth proteomics data analysis. p Find 3 features a swift open search workflow that is adept at uncovering less obvious information such as unexpected modifications or mutations that would have gone unnoticed using a conventional data analysis pipeline. In this protocol, we provide step-by-step instructions to help users mastering various types of data analysis using p Find 3 in conjunction with p Parse for data pre-processing and if needed, p Quant for quantitation. This streamlined p Parse-p Findp Quant workflow offers exceptional sensitivity, precision, and speed. It can be easily implemented in any laboratory in need of identifying peptides, proteins, or post-translational modifications, or of quantitation based on15N-labeling, SILAC-labeling, or TMT/i TRAQ labeling.

关键词： Mass spectrometry Protein identification Search engine Open-pFind quantitation

Non-target Screening and quantitation of Organic Chlorides in Oilfield Chemicals with Comprehensive Two Dimensional Gas Chromatography Coupled with Time of Flight Mass Spectrometry

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China Petroleum Processing & Petrochemical Technology 2017年第4期19卷 65-71页

作者： Zheng Kewen Guo Rui Ren Guofa Xia Lingyan Xiong Wei Zhang Jin Chu Xiaodong Yu Zhiqiang Institute of Environmental Pollution and Health School of Environmental and Chemical Engineering Shanghai University Shanghai 200072 SINOPEC Shengli Oilfield Branch Dongying 257000

Advanced analytical methodology based on comprehensive two-dimensional gas chromatography-time of flight mass spectrometry(GC×GC-TOFMS) is proposed for investigation of organic chlorides in different oilfield *** target screening was initially carried out on 8 suspected organic chlorides by evaluating the capability of the enhanced separation and reliable identification at a trace concentration. GC×GC-TOFMS allowed for the fast and automated analysis of organic chlorides at a level of 200 μg/L. This method was subsequently applied for non-target screening of organic chlorides in different oilfield chemicals at various locations across China. 22 organic chlorides were identified and verified by comparison with pure standards in the mixed sample. Finally, this method was used to determine the content of the organic chlorides in individual samples. The result showed that the organic chloride levels in 19 of the 39 tested oilfield chemicals were above the threshold limit of 1.0 mg/L.

关键词： GC×GC-TOFMS non-target screening oilfield chemicals organic chlorides quantitation

quantitation of ligupurpurosides B and C in rat plasma using HPLC-MS/MS

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Chinese Journal of Natural Medicines 2016年第6期14卷 473-480页

作者： CHEN Qian-Qian GUO Jian-Ru FENG Suo-Ming WANG Cai-Yun ZHANG Wei State Key Laboratory of Quality Research in Chinese Medicine Macao University of Science and Technology College of Pharmacy Xi'an Medial University

The present study was designed to develop a sensitive and selective specific high performance liquid chromatography(HPLC)-tandem mass spectrometric method(MS/MS) for the determination of ligupurpurosides B and C in rat plasma. The samples were prepared after protein precipitation and analyzed by liquid chromatography equipped with a C18 column interfaced with a triple quadrupole tandem mass spectrometer using ESI as the ionization source in the negative ion mode. The mobile phase consisted of water(0.01 % formic acid)–methanol(57 : 43, V/V) at the flow rate of 0.3 m L·min-1. The analytes and internal standard acteoside were both detected by use of multiple reaction monitoring mode. The total run time was 6.0 min. The method was linear in the concentration range of 2.5–500.0 ng·m L-1 and the lower limit of quantification(LLOQ) was 2.5 ng·m L-1. The intra-day and inter-day relative standard deviations across three validation runs over the entire concentration range were less than 9.8 %. The accuracy determined at three concentrations was within ± 6.1% in terms of relative error. In conclusion, this assay offers advantages in terms of expediency and suitability for the analysis of ligupurpuroside B and ligupurpuroside C in various biological fluids.

关键词： Ligupurpuroside B Ligupurpuroside C HPLC-MS/MS Pharmacokinetics quantitation

Ultra-High-Performance Liquid Chromatograph with Triple-Quadrupole Mass Spectrometer quantitation of Twelve Phenolic Components in Different Parts of Sarcandra glabra

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World Journal of Traditional Chinese Medicine 2021年第1期7卷 86-96页

作者： Jing-Guang Lu Cai-Yun Wang Da-Xin Chen Jing-Rong Wang Kai-Si Che Ming Zhong Wei Zhang Zhi-Hong Jiang State Key Laboratory of Quality Research in Chinese Medicines Macao Institute for Applied Research in Medicine and HealthMacao University of Science and TechnologyTaipaMacao Fujian Key Laboratory of Integrative Medicine on Geriatric Academy of Integrative MedicineFujian University of Traditional Chinese MedicineFuzhou Guangxi Key Laboratory of Traditional Chinese Medicine Quality Standards Guangxi Institute of Chinese Medicine and Pharmaceutical ScienceNanningPeople’s Republic of China

Objective:The study objective was to determine phenolic components for the quality control(QC)of cultivated Sarcandra glabra(Thunb.)Makino(***).Materials and Methods:A sensitive,ultra-high-performance liquid chromatography-tandem mass spectrometric method for the simultaneous determination of 12 phenolic components has been *** caffeoylquinic acids,two caffeoylshikimic acids,and four flavanonol glucosides were selected for the comprehensive analysis of distribution in different parts(root,stem,and leaf).Results:Twelve phenolic components were linear in the concentration range of 0.005–5.0μg/mL(R^(2)>0.995).The relative standard deviation of intra-day and inter-day precision across three validation runs over the entire concentration range was<5%.The recovery determined was within 5%in terms of relative *** results showed that the 12 phenolic compounds were mainly distributed in leaves and stems far more than those in ***:This study provided an ultra-high-performance liquid chromatograph with triple-quadrupole mass spectrometer quantitative method of 12 phenolic components for the QC of cultivated *** was found that the phenolic components were significantly accumulated in the aerial parts(stems and leaves)of cultivated ***.

关键词： Cultivation quantitation Sarcandra glabra ultra-high-performance liquid chromatograph with triple-quadrupole mass spectrometer

UHPLC-ESI-HRMS quantitation of Metabolites without Using Reference Standards: Impact of LC Flow Rate and Mobile Phase Composition on MS Responses

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American Journal of Analytical Chemistry 2013年第10期4卷 36-46页

作者： Sumithra Katragadda Samantha Mahmoud Dil M. Ramanathan Kean University New Jersey Center for Science Technology Education & Mathematics Union USA

During LC-MS quantitation of drugs for pharmacokinetic assessment, usually metabolites are not quantified due to the unavailability of reference standards. If a metabolite is quantified without a reference standard, then it is assumed that the LC-MS response to a drug is similar to that of its metabolite and the standard curve, of the parent compound, is used to quantitate the metabolite. This approach could result in an over or underestimation of the metabolite. To evaluate the impact of mobile phase composition on LC-MS response, the mobile phases were interchanged between methanol, acetonitrile and a 50/50 mixture of methanol/acetonitrile. UHPLC flow rates were varied from 200-500 μL/min, with and without the addition of reverse composition of mobile phases, at the parent drug retention time. This change was necessary to achieve uniform MS responses for drugs and their metabolites. In this study, HRMS data, obtained using orbi-trap, resulted in a linear response over a wider dynamic range than that obtained using the linear ion trap. Overall, the parameters, required for achieving standard free quantitation, are dependent upon the mobile phase composition and LC flow rates.

关键词： LC-MS Standard Free quantitation HRMS Pharmaceuticals Metabolites

Identification and quantitation of Cashmere (Pashmina) Fiber and Wool Using Novel Microchip Based Real-Time PCR Technology

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Journal of Textile Science and Technology 2018年第4期4卷 141-150页

作者： Rajwant Gill Sikander Gill Maxim Slyadnev Alexander Stroganov Lumex Instruments Canada Mission Canada

The textile industrial chain all over the world is facing a challenge of differentiating cashmere fiber from mixture of wool and other fibers in case cashmere stocks are adulterated with wool or other fibers. For identification of cashmere in such mixtures, the development of microchip based real-time PCR technology offers a very sensitive, specific, and accurate solution. The technology has been validated with cashmere and wool samples procured from distant farms, and from cashmere goats and sheep of different age and sex. Model samples with incremental raw cashmere or wool content were tested. The experimentally determined content was found to be comparable to the weighed content of the respective fibers in the samples. This technology may prove a cost cutter since it needs only 1.2 μl of the PCR reagent mix. It is substantially faster than traditional real-time PCR systems for being carried as miniature reaction volume in metal microchip. These features allow faster thermal equilibrium and thermal uniformity over the entire array of microreactors. For routine tests or in commercial set up, the microchips are available as ready-to-run with lyophilized reagents in its microreactors to which only 1 μl of the 10-fold diluted isolated DNA sample is added. The lyophilized microchips offer user-friendly handling in testing laboratories and help minimize human error.

关键词： Microchip Real-Time PCR Identification quantitation Cashmere Wool